The present disclosure relates to mutated LUX operon sequences and their use in producing autoluminescent glowing plants exhibiting improved light output.
Artificial and synthetic DNA sequences have gained extensive use with development of the field of biology in the past decade. The present disclosure relates to use of artificial nucleotide sequences in the field of bioluminescence, which is emission of light by living organisms. Bioluminescence of bacterial organisms is mediated by the bacterial LUX operon. The LUX operon encodes for the bacterial luciferase, the light emitting enzyme, as well as enzymes responsible for synthesis of luciferins, substrates required for the light emission reaction. The operon contains genes C-D-A-B-E(-G), where Lux A and Lux B code for the components of the luciferase and Lux C, D and E code for a fatty acid reductase complex producing an aldehyde necessary for the reaction. LuxG codes for an enzyme thought to participate in the turnover of the second luciferin, the flavin mononucleotide.
In biotechnology, genes of the LUX operon have a wide range of applications. For instance, the LUX operon is utilized as a reporter in a variety of bacterial and plant biosensors. Bacterial cells of naturally non-glowing species such as E. coli have been engineered to contain the LUX operon inducible by pre-determined classes of chemicals. These cells start glowing in the presence of these specific compounds, reporting on the composition or toxicity of the sample. Plants engineered with a fully functional LUX operon have been contemplated for use as phytosensors, monitoring the conditions of the plant and the environment.
In a further application of LUX technology, the present inventor developed the world's first autoluminescent glowing plants by employing genes of the LUX operon (Krichevsky et al. (2010) “Autoluminescent Plants”, PLoS One 12; 5(11)e15461; PCT International Publications WO 2009/017821 and WO 2011/106001). During the ensuing years, he has worked to improve the light output of the original autoluminescent plants, and has produced and successfully commercialized the first ornamental glowing plant varieties, demonstrating market interest in glowing plants produced via this technology.
The U.S. ornamentals market was sized at approximately $21 B in the early 2000's, and the entire worldwide market for ornamental plants has been estimated to be over $100 B.
The ornamental plant market is driven by innovation, where outdated varieties are inevitably replaced by new types of plants and flowers. New colors of roses and carnations, and new shapes and colors of petunias, find their way to the marketplace every year. Generation of new and esthetically pleasing varieties is known to be the key force driving the floriculture industry and stimulating its growth.
The inventor's previous application, U.S. Ser. No. 13/901,339, describes novel LUX operon sequences and mutations facilitating enhancement of light output compared to the use of native LUX operon sequences.
In view of the demand from consumers for even brighter glowing plants, further enhancements of LUX gene technology are needed.